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. 2013 Jan 31;8(1):e55724. doi: 10.1371/journal.pone.0055724

Figure 4. Akt activity is involved in VCP phosphorylation-induced protection against apoptosis and proliferation and cell division of AGS cells infected with H. pylori.

Figure 4

(A) AGS cells were transfected with control shRNA, Akt shRNA, or VCP shRNA for 36 hr, then were analyzed by immunoblotting. GAPDH served as the loading control. (B–D) Cells were stained with Annexin V/propidium iodide(PI) and the percentage of apoptotic cells analyzed by flow cytometry. (B) Apoptosis of AGS cells transfected with control shRNA, Akt shRNA, or VCP shRNA with or without H. pylori infection for 24 hr. **p<0.01 relative to cells uninfected with H. pylori. § p<0.01 relative to cells transfected with control shRNA. Data are expressed as means ± SD. (C) Apoptosis of AGS cells with or without pretreatment with 10 µM LY294002 for 3 hr with or without H. pylori infection for 24 hr. **p<0.01 relative to cells uninfected with H. pylori. § p<0.01 relative to cells treated with DMSO. Data are expressed as means ± SD. (D) Apoptosis of AGS cells transfected withVCP-Flag or Tri-mut VCP-Flag with or without H. pylori infection for 24 hr. (E) Proliferation of AGS cells transfected with VCP-Flag or Tri-mut VCP-Flag with or without H. pylori infection for 24 hr. Cell proliferation was measured using the MTS assay. (F) Percentage of cells in G0/G1 phase in AGS cells transfected with VCP-Flag or Tri-mut VCP-Flag with or without H. pylori infection for 24 hr. The percentage of cells in G0/G1 was examined by flow cytometry. In D–F, *p<0.05, **p<0.01 relative to cells uninfected with H. pylori. § p<0.01 relative to cells transfected with WT VCP-Flag. Data are expressed as means ± SD.